+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-14469 | |||||||||||||||||||||
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タイトル | Structure of yeast RNA Polymerase III-Ty1 integrase complex at 2.9 A (focus subunit C11 terminal Zn-ribbon in the funnel pore). | |||||||||||||||||||||
マップデータ | Cryo-EM structure of the crosslinked RNA Polymerase III-Ty1 integrase complex at 2.9 (focus subunit C11 C-terminal Zn-ribbon in the funnel pore). | |||||||||||||||||||||
試料 |
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キーワード | targeted DNA integration / Ty1 retrotransposon / Ty1 integrase / RNA 19 polymerase III / transcription | |||||||||||||||||||||
機能・相同性 | 機能・相同性情報 transposition / ribonuclease H / RNA Polymerase I Transcription Initiation / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / mRNA Capping / RNA polymerase II transcribes snRNA genes / TP53 Regulates Transcription of DNA Repair Genes ...transposition / ribonuclease H / RNA Polymerase I Transcription Initiation / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / mRNA Capping / RNA polymerase II transcribes snRNA genes / TP53 Regulates Transcription of DNA Repair Genes / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Initiation And Promoter Clearance / 加水分解酵素; プロテアーゼ; ペプチド結合加水分解酵素; アスパラギン酸プロテアーゼ / RNA-templated transcription / RNA Polymerase II Pre-transcription Events / termination of RNA polymerase III transcription / Formation of TC-NER Pre-Incision Complex / RNA polymerase III activity / transcription initiation at RNA polymerase III promoter / termination of RNA polymerase I transcription / RNA Polymerase I Promoter Escape / nucleolar large rRNA transcription by RNA polymerase I / Gap-filling DNA repair synthesis and ligation in TC-NER / transcription by RNA polymerase I / transcription initiation at RNA polymerase I promoter / Estrogen-dependent gene expression / transcription by RNA polymerase III / Dual incision in TC-NER / transcription elongation by RNA polymerase I / tRNA transcription by RNA polymerase III / RNA polymerase I activity / RNA polymerase I complex / RNA polymerase III complex / RNA polymerase II, core complex / nucleotidyltransferase activity / transcription initiation at RNA polymerase II promoter / transcription elongation by RNA polymerase II / ribonucleoside binding / DNA integration / DNA-directed 5'-3' RNA polymerase activity / RNA-directed DNA polymerase / DNA-directed RNA polymerase / RNA-directed DNA polymerase activity / RNA-DNA hybrid ribonuclease activity / peroxisome / ribosome biogenesis / single-stranded DNA binding / DNA recombination / transcription by RNA polymerase II / nucleic acid binding / DNA-directed DNA polymerase / aspartic-type endopeptidase activity / DNA-directed DNA polymerase activity / protein dimerization activity / viral translational frameshifting / nucleotide binding / nucleolus / mitochondrion / proteolysis / DNA binding / RNA binding / zinc ion binding / nucleoplasm / ATP binding / nucleus / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||||||||||||||
生物種 | Saccharomyces cerevisiae S288C (パン酵母) | |||||||||||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.9 Å | |||||||||||||||||||||
データ登録者 | Nguyen PQ / Huecas S / Plaza-Pegueroles A / Fernandez-Tornero C | |||||||||||||||||||||
資金援助 | European Union, フランス, スペイン, 6件
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引用 | ジャーナル: Nat Commun / 年: 2023 タイトル: Structural basis of Ty1 integrase tethering to RNA polymerase III for targeted retrotransposon integration. 著者: Phong Quoc Nguyen / Sonia Huecas / Amna Asif-Laidin / Adrián Plaza-Pegueroles / Beatrice Capuzzi / Noé Palmic / Christine Conesa / Joël Acker / Juan Reguera / Pascale Lesage / Carlos Fernández-Tornero / 要旨: The yeast Ty1 retrotransposon integrates upstream of genes transcribed by RNA polymerase III (Pol III). Specificity of integration is mediated by an interaction between the Ty1 integrase (IN1) and ...The yeast Ty1 retrotransposon integrates upstream of genes transcribed by RNA polymerase III (Pol III). Specificity of integration is mediated by an interaction between the Ty1 integrase (IN1) and Pol III, currently uncharacterized at the atomic level. We report cryo-EM structures of Pol III in complex with IN1, revealing a 16-residue segment at the IN1 C-terminus that contacts Pol III subunits AC40 and AC19, an interaction that we validate by in vivo mutational analysis. Binding to IN1 associates with allosteric changes in Pol III that may affect its transcriptional activity. The C-terminal domain of subunit C11, involved in RNA cleavage, inserts into the Pol III funnel pore, providing evidence for a two-metal mechanism during RNA cleavage. Additionally, ordering next to C11 of an N-terminal portion from subunit C53 may explain the connection between these subunits during termination and reinitiation. Deletion of the C53 N-terminal region leads to reduced chromatin association of Pol III and IN1, and a major fall in Ty1 integration events. Our data support a model in which IN1 binding induces a Pol III configuration that may favor its retention on chromatin, thereby improving the likelihood of Ty1 integration. | |||||||||||||||||||||
履歴 |
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-構造の表示
添付画像 |
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-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_14469.map.gz | 96.6 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-14469-v30.xml emd-14469.xml | 43.5 KB 43.5 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_14469_fsc.xml | 10.6 KB | 表示 | FSCデータファイル |
画像 | emd_14469.png | 30.4 KB | ||
Filedesc metadata | emd-14469.cif.gz | 11.2 KB | ||
その他 | emd_14469_half_map_1.map.gz emd_14469_half_map_2.map.gz | 80.8 MB 80.8 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-14469 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-14469 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_14469_validation.pdf.gz | 1.1 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_14469_full_validation.pdf.gz | 1.1 MB | 表示 | |
XML形式データ | emd_14469_validation.xml.gz | 17.8 KB | 表示 | |
CIF形式データ | emd_14469_validation.cif.gz | 23.5 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-14469 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-14469 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_14469.map.gz / 形式: CCP4 / 大きさ: 103 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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注釈 | Cryo-EM structure of the crosslinked RNA Polymerase III-Ty1 integrase complex at 2.9 (focus subunit C11 C-terminal Zn-ribbon in the funnel pore). | ||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.06 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-ハーフマップ: Cryo-EM half-map2 of the crosslinked RNA Polymerase III-Ty1...
ファイル | emd_14469_half_map_1.map | ||||||||||||
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注釈 | Cryo-EM half-map2 of the crosslinked RNA Polymerase III-Ty1 integrase complex at 2.9 (focus subunit C11 C-terminal Zn-ribbon in the funnel pore). | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: Cryo-EM half-map1 of the crosslinked RNA Polymerase III-Ty1...
ファイル | emd_14469_half_map_2.map | ||||||||||||
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注釈 | Cryo-EM half-map1 of the crosslinked RNA Polymerase III-Ty1 integrase complex at 2.9 (focus subunit C11 C-terminal Zn-ribbon in the funnel pore). | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Cryo-EM map of the crosslinked RNA Polymerase III-Ty1 integrase c...
+超分子 #1: Cryo-EM map of the crosslinked RNA Polymerase III-Ty1 integrase c...
+分子 #1: DNA-directed RNA polymerase III subunit RPC1
+分子 #2: DNA-directed RNA polymerase III subunit RPC2
+分子 #3: DNA-directed RNA polymerases I and III subunit RPAC1
+分子 #4: DNA-directed RNA polymerase III subunit RPC9
+分子 #5: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+分子 #6: DNA-directed RNA polymerases I, II, and III subunit RPABC2
+分子 #7: DNA-directed RNA polymerase III subunit RPC8
+分子 #8: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+分子 #9: DNA-directed RNA polymerase III subunit RPC10
+分子 #10: DNA-directed RNA polymerases I, II, and III subunit RPABC5
+分子 #11: DNA-directed RNA polymerases I and III subunit RPAC2
+分子 #12: DNA-directed RNA polymerases I, II, and III subunit RPABC4
+分子 #13: DNA-directed RNA polymerase III subunit RPC5
+分子 #14: DNA-directed RNA polymerase III subunit RPC4
+分子 #15: DNA-directed RNA polymerase III subunit RPC3
+分子 #16: DNA-directed RNA polymerase III subunit RPC6
+分子 #17: DNA-directed RNA polymerase III subunit RPC7
+分子 #18: Integrase
+分子 #19: Unknown RNA Polymerase III chain
+分子 #20: ZINC ION
+分子 #21: MAGNESIUM ION
+分子 #22: water
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
緩衝液 | pH: 7.5 |
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凍結 | 凍結剤: ETHANE |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 42.45 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD 最大 デフォーカス(公称値): 2.8000000000000003 µm 最小 デフォーカス(公称値): 1.0 µm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |